title: Quantification of matrix metalloproteinases and tissue inhibitors of metalloproteinase in prostatic tissue: analytical aspects.
abstract: BACKGROUND
  The balance between matrix metalloproteinases (MMP) and the tissue inhibitors of metalloproteinases (TIMP) has been seen as important during tumor invasion and progression. The determination of these components needs a special strategy of tissue preparation. This analytical problem has not been considered for prostatic tissue.
  
  
  METHODS
  We adapted an extraction method consisting of two extraction steps with 0.25% Triton X-100/CaCl2 solution and two heat extraction steps at 60 degrees C for 4 min. This combination allowed a complete extraction of MMP (measured as enzyme activity) and TIMP-1 (measured with an ELISA test) from cancerous and normal prostatic tissue samples.
  
  
  RESULTS
  The median values for cancerous vs. normal MMPs (50.8 mU/g wet tissue and 1,580 mU/g protein vs. 88.8 and 2,497) and TIMP-1 (4.49 micrograms/g wet tissue and 96.7 micrograms/g protein vs. 12.4 and 237.8) were significantly lower, whereas the respective ratios for MMP/ TIMP-1 (11.1 vs. 4.0 on wet weight and 15.5 vs. 5.3 on protein basis) were significantly higher.
  
  
  CONCLUSIONS
  An optimized extraction procedure was elaborated for determining MMPs and TIMP-1 in prostatic tissue samples. The increased ratio of MMP/TIMP-1 can be interpreted as an indicator of the imbalance between MMP and TIMP, characteristic of prostate carcinoma tissue.
authors.affiliation: Department of Urology, University Hospital Charité, Humboldt University, Berlin, Germany. jung@rz.charite.hu-berlin.de
authors.forename: K
authors.initials: K
authors.lastname: Jung
authors.forename: M
authors.initials: M
authors.lastname: Lein
authors.forename: N
authors.initials: N
authors.lastname: Ulbrich
authors.forename: B
authors.initials: B
authors.lastname: Rudolph
authors.forename: W
authors.initials: W
authors.lastname: Henke
authors.forename: D
authors.initials: D
authors.lastname: Schnorr
authors.forename: S A
authors.initials: SA
authors.lastname: Loening
mesh_terms: D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D006801:Humans; D008297:Male; D008666:Metalloendopeptidases / Q000032:analysis*; D008875:Middle Aged; D011467:Prostate / Q000737:chemistry* / Q000166:cytology; D011471:Prostatic Neoplasms / Q000737:chemistry* / Q000473:pathology; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000032:analysis; D019714:Tissue Inhibitor of Metalloproteinases / Q000032:analysis*
chemical_list: D019715:Tissue Inhibitor of Metalloproteinase-1; D019714:Tissue Inhibitor of Metalloproteinases; D008666:Metalloendopeptidases